Stefano Sartori (Presenter)
Eureka srl Lab Division
Authorship: E.BASSOTTI, V. CONIGLIONE, M. MATTEUCCI et S. SARTORI
Eureka srl Lab Division
| Short Abstract Therapeutic drug monitoring (TDM) plays a key role in understanding what is the most effective therapy to be applied to the patient and, at the same time, putting in the foreground his state of health. This work describes results obtained using Eureka Lab Division ready to use kit. The kit permits to dose in a single chromatographic run and with single extraction procedure, 9 most common antibiotic drugs in human plasma by LC-MS/MS in a sensitive and accurate way. Its easiness of use and its robustness match perfectly with the needs of the laboratory routine. |
Long Abstract
Eureka Lab Division Antibiotic Drugs in human plasma ready to use kit permits the simultaneous analysis of the most important antibiotics (Daptomycin, Vancomycin, Streptomycin, Linezolid, Levofloxacin, Ciprofloxacin, Gentamicin, Amikacin, Teicoplanin) in a single chromatographic run after a simple matrix deproteinization.
To analyse correctly the full panel in routine it's necessary to have almost a medium level mass QQQ detector coupled with an HPLC or UHPLC.
5 levels of plasma calibrators and 2 levels of plasma controls are available, covering clinical range concentration values.
Plasma matrix sample is deproteinized with an appropriate reagent containing Internal Standards, centrifuged, diluted and injected into LC-MS/MS.
This method is simple, robust, sensitive and fast.
Chromatographic run is 5-6 minutes length.
Calibration curve is on 5 concentration levels and is manufactured to cover clinical therapeutic range, usually less than the lower limit and more than the higher level. Controls are on 2 levels, low and high concentrations.
Calibrators and Controls are lyophilized human plasma.
All analytes are simply detected, using same column and same reagents, with a low level LC-MS/MS except for Gentamicin and Amikacin that are detected correctly, at low concentration, with a medium level mass spectrometer.
The LOQ obtained using a medium level LC-MS/MS is 0,5 mg/l for Daptomycin, Linezolid, Levofloxacin, Ciprofloxacin, Streptomycin and Teicoplanin , and 1 mg/l for Gentamicin , Vancomycin and Amikacin.
The average of CV% INTRA-serie of all analytes is 6.4%, and 10,5% is the CV% INTER-serie average.
Features like simplicity, robustness and reproducibility obtained with this method make it a suitable kit to support the clinical laboratory routine.
References & Acknowledgements:
[1] Nobuhito Shibata, Makoto Ishida, Yarasani Venkata Rama Prasad, Weihua Gao, Yukako Yoshikawa, Kanji Takada, Highly sensitive quantification of vancomycin in plasma samples using liquid chromatography–tandem mass spectrometry and oral bioavailability in rats, Journal of Chromatography B, 789 (2003) 211–218
[2] Chang Cheng, Shaorong Liu, Deqing Xiao, John Hollembaek, Lili Yao, Jing Lin, Steven Hansel, LC–MS/MS method development and validation for the determination of polymyxins and vancomycin in rat plasma, Journal of Chromatography B, 878 (2010) 2831–2838.
[3] David N. Heller, James O. Peggins, Cristina B. Nochetto, Michelle L. Smith, O. Alberto Chiesa, Keesla Moulton, LC/MS/MS measurement of gentamicin in bovine plasma, urine, milk, and biopsy samples taken from kidneys of standing animals, Journal of Chromatography B, 821 (2005) 22–30
[4] Josè Diana, Dora Visky, Eugène Roets, Jos Hoogmartens, Development and validation of an improved method for the analysis of vancomycin by liquid chromatography selectivity of reversed-phase columns towards vancomycin components, Journal of Chromatography A, 996 (2003) 115–131.
[5] Wei-xia Zhu, Ji-zhou Yanga,WeiWei , Ya-feng Liu, Shu-sheng Zhang, Simultaneous determination of 13 aminoglycoside residues in foods of animal origin by liquid chromatography–electrospray ionization tandem mass spectrometry with two consecutive solid-phase extraction steps, Journal of Chromatography A 1207 (2008) 29–37
[6] Sara Bogialli, Roberta Curini, Antonio Di Corcia, Aldo Laganà, Monica Mele, Manuela Nazzari, Simple confirmatory assay for analyzing residues of aminoglycoside antibiotics in bovine milk: hot water extraction followed by liquid chromatography -tandem mass spectrometry, Journal of Chromatography A, 1067 (2005) 93–100
[7] http://host.uniroma3.it/centri/cisdic/laboratorio_chimica_organica/lezioni%5CL0 .pdf
[8] Douglas A. Skoog, James F. Holler, Stanley R. Crouch, Chimica analitica strumentale, II edizione, 2009
[9] xxx, basic of LC/MS, Agilent technologies, (2010).
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